Background/Aims: This study aimed to explore the effect of gut microbiota-regulated Kupffer cells (KCs) on colorectal cancer (CRC) liver metastasis. Methods: A series of in vivo and in vitro researches were showed to demonstrate the gut microbiota and its possible mechanism in CRC liver metastasis. Results: Fewer liver metastases were identified in the ampicillin-streptomycin-colistin and colistin groups. Increased proportions of Parabacteroides goldsteinii, Bacteroides vulgatus, Bacteroides thetaiotaomicron, and Bacteroides uniforms were observed in the colistin group. The significant expansion of KCs was identified in the ampicillin-streptomycin-colistin and colistin groups. B.vulgatus levels were positively correlated with KC levels. More liver metastases were observed in the vancomycin group. An increased abundance of Parabacteroides distasonis and Proteus mirabilis and an obvious reduction of KCs were noted in the vancomycin group. P. mirabilis levels were negatively related to KC levels. The number of liver metastatic nodules was increased in the P. mirabilis group and decreased in the B. vulgatus group. The number of KCs decreased in the P. mirabilis group and increased in the B. vulgatus group. In vitro, as P. mirabilis or B. vulgatus doses increased, there was an opposite effect on KC proliferation in dose- and time-dependent manners. P. mirabilis induced CT26 cell migration by controlling KC proliferation, whereas B. vulgatus prevented this migration. Conclusions An increased abundance of P. mirabilis and decreased amount of B. vulgatus play key roles in CRC liver metastasis, which might be related to KC reductions in the liver.

Objective: To evaluate the differences of serum total light chain(sTLC), urine total light chain(uTLC) and serum free light chain(sFLC) in different stages of chronic kidney disease(CKD) and their correlation with renal function indexes. To investigate the predictive value of light chain indexes in CKD staging. Methods: 292 patients with CKD were analyzed retrospectively, and plasma cell diseases, acute kidney injury and tumor diseases were excluded. According to the estimated glomerular filtration rate(eGFR), CKD patients were divided into five groups from CKD 1 stage to CKD 5 stage. The levels of sTLC, uTLC, sFLC and corresponding biochemical indexes of CKD patients were detected, and the differences and correlations among the indexes of each group were compared. The receiver operating curve(ROC curve) was used to analyze the predictive value of each light chain index in CKD stage, with CKD1-2 stage combined as control group and CKD3-5 stage combined as case group. Results: There was no significant difference in sTLC κ, sTLC λ, sTLC κ/λ and sFLC κ/λ among CKD1-5 stage(P>0.05). There were significant differences between sFLC κ, sFLC λ and uTLC κ, uTLC λ among CKD1-5 stage(P<0.05), which increased with the increase of CKD staging. The correlation between sFLC κ, sFLC λ and serum creatinine(Scr), blood urea nitrogen(BUN), eGFR were better than uTLC κ, uTLC λ(P<0.001). The sTLC κ, sTLC λ, sTLC κ/λ and sFLC κ/λ had no correlation with renal function indexes(P>0.05). The best critical points of sFLC κ and sFLC λ for predicting CKD3-5 stage were 35.4 mg/L and 52.8 mg/L, and AUC was 0.916(0.883-0.949) and 0.915(0.881-0.949), which were higher than uTLC κ and uTLC λ,AUC was 0.811(0.754-0.869) and 0.787(0.728-0.846), respectively. Conclusion With the increase of CKD staging, the levels of sFLC and uTLC gradually increase. The sFLC and uTLC can effectively predict patients with CKD3 and above, which has an important reference value in stratified management of patients with CKD.

In recent years, more and more attention has been paid to the integration of nano-material graphene and field effect transistor to construct biosensor for detecting biomolecules.This is mainly due to the following advantages of graphene field effect transistor inculding high sensitivity and specificity, rapid analysis, label-free detection, cheap price, miniaturization and integration.This review summarizes the structure and working principle of graphene field effect transistor biosensor, the preparation and functionalization of graphene, and the application of graphene field effect transistor in medical detections.

Objective: To evaluate the effect of Annexin-A1 mimetic peptide Ac2-26 on activation of astrocytes in rats. Methods: The primarily cultured astrocytes from the cortex of fetal Sprague-Dawley rats after 4 passages were divined into 4 groups (n=24 each) using a random number table method: control group (C group), LPS group, LPS+ scramble peptide group (LPS+ Src group) and LPS+ Ac2-26 group.LPS was added to LPS group with the final concentration of 1 mg/ml.LPS at the final concentration of 1 mg/ml and scramble peptide at the final concentration of 3.3 mmol/L were added to LPS+ Src group.LPS at the final concentration of 1 mg/ml and Ac2-26 at the final concentration of 3.3 mmol/L were added to LPS+ Ac2-26 group.After 24-h incubation, the cell survival rate was measured by CCK-8 assay, the migration was determined by Transwell assay, the concentrations of tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β), monocyte chemoattractant protein-1 (MCP-1) and macrophage inflammatory protein-1a (MIP-1a) in the supernatant were measured (by enzyme-linked immunosorbent assay), and the expression of glial fibrillary acidic protein (GFAP), extracellular signal-regulated kinase (ERK), phosphorylated ERK (p-ERK), c-Jun N-terminal kinase (JNK), phosphorylated JNK (p-JNK), p38 mitogen-activated protein kinase (p38MAPK), and phosphorylated p38MAPK (p-p38MAPK) in astrocytes was detected by Western blot. Results: Compared with group C, the expression of GFAP was significantly up-regulated, and the cell mobility, concentrations of TNF-α, IL-1β, MCP-1 and MIP-1α in the supernatant, p-ERK/ERK ratio, p-JNK/JNK ratio and p-p38MAPK/p38MAPK ratio were increased (P<0.05), and no significant change was found in the cell survival rate in group LPS (P>0.05). Compared with group LPS, the expression of GFAP was significantly down-regulated, and the cell mobility, concentrations of TNF-α, IL-1β, MCP-1 and MIP-1α in the supernatant, p-JNK/JNK ratio and p-p38MAPK/p38MAPK ratio were decreased in group LPS+ Ac2-26 (P<0.05), and no significant change was found in the parameters mentioned above in group LPS+ Src (P>0.05). Conclusion Ac2-26 can inhibit activation of astrocytes and produces anti-inflammatory effect in rats.

Objective To explore the expression of tiny RNA-25 (microRNA-25, miR-25) in the plasma、tissues of triple-negative breast cancer(TNBC) patients and cell lines, to investigate the potential molecular mechanisms of miR-25 on migration and invasion of TNBC. Methods Real-time fluorescent quantitative PCR was used to detect the expression of miR-25 in the plasma of TNBC patients. Linked omics web platform was used to analyse miR-25 level in samples of TNBC and non-TNBC. Real-time fluorescent quantitative PCR was also used to detect the miR-25 level in TNBC cell lines. The wound healing and transwell assay was applied to assess the effects on migration and invasion of TNBC cell lines which transfected with miR-25 inhibitor or the negative control. The luciferase reporter assay was used to validate the relationship between miR-25 and the sphingosine-1-phosphate phosphatase 1 (SGPP1) in HEK293T cell. The wound healing and transwell assay was used to detect the migration and invasion ability of TNBC cell lines when cotransfected with pCMV6-SGPP1 and miR-25. Furthermore, Western blot was performed to detect the SGPP1 level in TNBC cell lines. Results The expression of miR-25 was significantly elevated in the plasma of 86 TNBC patients compared with the healthy controls (P value was 0.031). LinkedOmics web platform analysis showed that miR-25 expression was significantly higher in TNBC samples than in non-TNBC samples with Luminal A or Luminal B (P value was<0.001 and 0.006). The level of miR-25 was also elevated in TNBC cell lines HS578T, HCC1806, MDA-MB-231 and BT549(P value was 0.006, 0.01, 0.029 and 0.046). The MDA-MB-231 and HS578T cells which transfected with miR-25 inhibitor exhibited a significant slower wound healing rate than control (P value was 0.035 and 0.001). At the same time, when transfected with miR-25 inhibitor, MDA-MB-231 and HS578T both exhibited a decreased invasion ability compared with the control group(P value was 0.002 and 0.001). LinkedOmics web platform analysis showed that sphingosine-1-phosphate phosphatase 1 (SGPP1) gene level was negatively correlated with miR-25 in the tissues of TNBC patients (P value was 0.037). The luciferase reporter assay validated that SGPP1 was a directed target of miR-25. The western blot assay indicated that the SGPP1 level was increased in MDA-MB-231 and HS578T after transfection with miR-25 inhibitor. Over-expression of SGPP1 could abrogate the positive effects of miR-25 on migration and invasion when pCMV6-SGPP1 was cotransfected with miR-25 (P value was all 0.002). Conclusions MiR-25 was elevated in both plasma and tissues of TNBC patients and also increased in TNBC cell lines. Transfection of MDA-MB-231 and HS578T cells with miR-25 inhibitor resulted in reduced migration and invasion. Moreover, SGPP1 was identified as a novel target of miR-25. The ability of miR-25 to promote TNBC cell migration and invasion is attributable to its effect on SGPP1 suppression.

Objective To evaluate the efficacy of cardiac output ( CO ) and corrected flow time ( FTc) monitored by ultrasonic cardiac output monitor ( USCOM) in guiding volume therapy in patients un-dergoing laparoscopic colorectal surgery. Methods Eighty American Society of Anesthesiology physical sta-tus Ⅰ or Ⅱ patients, aged 18-60 yr, with body mass index of 18-25 kg∕m2 , undergoing laparoscopic colorectal surgery, were divided into 2 groups ( n=40 each) using a random number table method: control group ( group C ) and USCOM-guided fluid therapy group ( group U ) . Mean arterial pressure was main-tained at 60-100 mmHg, central venous pressure at 5-10 cmH2 O and urine volume>0. 5 ml·kg-1 ·h-1 u-sing conventional fluid therapy in group C. In group U, goal-directed fluid therapy was performed under the monitoring of USCOM, FTc was maintained at 326-400 ms and CO at 4. 2-5. 9 L∕min. The volume of crys-talloid and colloid solution, total volume of fluid infused, blood loss, urine volume and requirement for va-soactive agents during surgery and time of surgery were recorded. Blood samples were collected from the left radial artery for determination of the blood lactate concentration immediately after anesthesia induction, at the end of the operation and at 1 and 2 days after surgery. The time of passing the first flatus after surgery, adverse cardiovascular events ( hypertension, hypotension, cardiac insufficiency ) , pulmonary complica-tions ( pulmonary edema, pulmonary atelectasis) , and oliguria and anuria within 24 h after surgery, and length of hospital stay were recorded. Results Compared with group C, the volume of crystalloid, total volume of fluid infused and urine volume were significantly decreased during surgery, the volume of colloid solution was increased during surgery, the blood lactate concentration was decreased at the end of surgery, the incidence of postoperative cardiovascular and pulmonary complications was decreased, and the time of passing the first flatus after surgery and length of hospital stay were shortened in group U ( P<0. 05) . Con-clusion USCOM-monitored CO and FTc produces better efficacy in guiding volume therapy and is helpful for improving recovery in patients undergoing laparoscopic colorectal surgery.

Objective To investigate the role of OMA1 in acute kidney injury (AKI) induced by lipopolysaccharide (LPS).Methods OMA1 wild-type and knocked out mice (8 week old) were injected with 10 mg/kg body weight of LPS.The model was confirmed by testing mouse serum creatinine and blood urea nitrogen.The apoptosis in mouse kidney cortex was examined by TUNEL staining and cleaved caspase 3.In vitro,in humam kidney proximal tubular cells (HK2) were knocked down OMA1 by transfecting OMA1 shRNA,with the scramble shRNA being used as negative control of transfection.HK2 cells were cultured with 5 μg/ml of LPS for 24 hours to induce apoptosis.DAPI staining of cells and caspase-3 activity were applied to test apoptosis.The images of mitochondria in cells were obtained by transfection of mito-green plasmid and OMA1 shRNA.Western blotting was used to exam the OMA1 and Cytochrome C expressions.Resudts Compared with OMA1 KO mice,LPS induced more severe AKI of WT mice with higher Scr [(97.2±26.5) μmol/L vs (53.0±17.7) μmol/L,P ＜ 0.05] and BUN [(43.3± 13.7) mmol/L vs (29.7±7.7) mmol/L,P ＜ 0.05].Moreover,there were more apoptosis cells in kidney cortex in WT mice than in OMA1 KO mice [(75.4± 26.1)/ram2 vs (38.3± 14.4)/mm2,P＜ 0.05].About 46％ of OMA1 expressions in HK2 cells were inhibited by OMA1 shRNA transfection (P ＜ 0.05).Further,OMA1 shRNA cells with LPS stimulation had decreased mitochondria fragmentation [(29.8±10.9)％ vs (43.2±6.8)％,P ＜ 0.05],Cytochrome C release [(37.0±12.3)％ vs (76.0±26.2)％,P ＜ 0.05],and cell apoptosis [(13.2±3.9)％ vs (25.0±7.1)％,P ＜ 0.05] as compared with control cells.Conclusion Knockdown of OMA1 alleviated septic AKI through inhibition of cell apoptosis,mitochondria fragmentation,and Cytochrome C release.

Objective To investigate the expression of anti-endothelial cell antibody AECA in the emphysema rats induced by smoking and to analyze the intervention effect of methylprednisolone on it.Methods Thirty-nine rats were randomly divided into the control group,smoking rat emphysema model group (model group) and methylprednisolone intervention group (intervention group).The model group and intervention group conducted the 1-month passive smoking by smog exposure.The intervention group was intraperitoneally injected by methylprednisolone(once daily,6 d per week).After exposing to smog for 90 d,the differences of serum AECA level,IL-8,MMP-9 and TNF-α level in bronchoalveolar lavage fluid(BALF),lung MLI and mean alveolar number (MAN) were compared among groups.Results Compared with the control group and theintervention group,the level of serum AECA in the model group was significantly increased (P＜0.05);Compared with the control group and intervention group,the IL-8,TNF-α and MMP-9 levels of BALF in the model group were also increased(P＜0.05).Conclusion ACEA participates in the smoking induced emphysema formation;methylprednisolone may decrease the level of AECA and cell inflammatory factors and affects the emphysema formation.

Objective To investigate the effects of Annexin-A1 ( Anxa1 ) gene silencing induced by siRNA on the growth and migration of microglial BV-2 cells and its possible mechanisms.Methods A synthesized siRNA duplex targeting Anxa1 gene was transfected into BV-2 cells.The efficiency of siRNA-in-duced Anxa1 gene silencing was evaluated on both mRNA and protein levels by using reverse-transcription PCR and Western blot assay.MTT assay was performed to measure the proliferation of BV-2 cells with si-lenced expression of Anxa1 gene.Flow cytometry with Annexin V-FITC/PI double staining was used to de-tect the apoptosis rate of BV-2 cells.Transwell chambers were used to analyze the effects of siRNA-induced Anxa1 gene silencing on the migration of BV-2 cells.Western blot assay was performed to detect the expres-sion of signaling proteins related to cell cycle and migration.Results Compared with the siRNA negative control ( siRNA-NC) group, the inhibitory rates of siRNA-induced Anxa1 gene silencing on the proliferation of BV-2 cells were significantly increased at the time points of 24 h, 48 h and 72 h after intervention [(16.9 ±2.1)%, (23.1±3.6)%and (42.4±1.7)%vs (1.35±0.5)%, (2.06±0.7)% and (8.65±0.9)%, P<0.05 ].The apoptosis rate of BV-2 cells transfected with Anxa1 siRNA was (18.4±2.1)%, which was significantly elevated as compared with that of the siRNA-NC group (5.2±0.3)%and control group (4.3±0.2)%.Cell migration of the Anxa1 siRNA transfected BV-2 cells was inhibited remarkably at 48 h as com-pared with that of the siRNA-NC group (28.7±5.2 vs 173.4±11.4, P<0.01).Moreover, the suppressed expression of Cyclin D1 protein and activation of p38 and JNK signaling pathways were induced by silenced expression of Anxa1 gene in BV-2 cells.Conclusion The growth and migration of BV-2 cells were signifi-cantly inhibited by silencing the expression of Anxa1 gene with siRNA, the possible mechanisms might be associated with the suppressed expression of Cyclin D1protein and the activation of p38 and JNK signaling pathways.

The recent explosive outbreak of Zika virus (ZIKV) infection has been reported in South and Central America and the Caribbean. Neonatal microcephaly associated with ZIKV infection has already caused a public health emergency of international concern. No specific vaccines or drugs are currently available to treat ZIKV infection. The ZIKV helicase, which plays a pivotal role in viral RNA replication, is an attractive target for therapy. We determined the crystal structures of ZIKV helicase-ATP-Mn(2+) and ZIKV helicase-RNA. This is the first structure of any flavivirus helicase bound to ATP. Comparisons with related flavivirus helicases have shown that although the critical P-loop in the active site has variable conformations among different species, it adopts an identical mode to recognize ATP/Mn(2+). The structure of ZIKV helicase-RNA has revealed that upon RNA binding, rotations of the motor domains can cause significant conformational changes. Strikingly, although ZIKV and dengue virus (DENV) apo-helicases share conserved residues for RNA binding, their different manners of motor domain rotations result in distinct individual modes for RNA recognition. It suggests that flavivirus helicases could have evolved a conserved engine to convert chemical energy from nucleoside triphosphate to mechanical energy for RNA unwinding, but different motor domain rotations result in variable RNA recognition modes to adapt to individual viral replication.
Crystallography, X-Ray ; Protein Domains ; RNA Helicases ; chemistry ; RNA, Viral ; chemistry ; Viral Proteins ; chemistry ; Zika Virus ; enzymology